Development of a reverse transcription-droplet digital PCR method for absolute quantification of citrus tatter leaf virus

被引：2
|
作者：
Zhao, Jinfa [1 ]
Wang, Ying [1 ]
Zhang, Xingkai [1 ]
Zeng, Ting [1 ]
Qin, Yangyang [1 ]
Shen, Wanxia [1 ]
Zhang, Wei [1 ,2 ]
Zhou, Yan [1 ]
机构：
[1] Southwest Univ, Citrus Res Inst, Natl Citrus Engn & Technol Res Ctr, Chongqing 400712, Peoples R China
[2] Agr & Rural Affairs Sichuan Prov, Plant Protect Stn Dept, Chengdu 640041, Peoples R China
来源：
EUROPEAN JOURNAL OF PLANT PATHOLOGY | 2021年 / 161卷 / 03期
基金：
国家重点研发计划;
关键词：
citrus tatter leaf virus; Droplet digital PCR; Quantitation; ASSAY; APPLE;
D O I：
10.1007/s10658-021-02357-x
中图分类号：
S3 [农学（农艺学）];
学科分类号：
0901 ;
摘要：
Tatter leaf disease caused by citrus tatter leaf virus (CTLV) is a serious threat to the citrus industry. In this study, a reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR) assay for detection of CTLV was established by optimizing the reaction conditions. The dynamic range of the detection of CTLV RNA transcripts by RT-ddPCR was from 5 to 5 x 10(5) copies/mu L, and the sensitivity was 10 times higher than that of RT real-time PCR. Both techniques showed a high degree of linearity and quantitative correlation. The number of positive samples that was identified in field citrus samples using RT-ddPCR (62.3%) was higher than that of RT-qPCR (49.2%). The results demonstrated that the RT-ddPCR assay is a promising tool for improving CTLV detection.
引用
收藏
页码：743 / 749
页数：7
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